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  • 99
    Thermo Fisher pfa
    Pfa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10997 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa/product/Thermo Fisher
    Average 99 stars, based on 10997 article reviews
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    pfa - by Bioz Stars, 2020-11
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    99
    Millipore pfa
    Pfa, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 16222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa/product/Millipore
    Average 99 stars, based on 16222 article reviews
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    pfa - by Bioz Stars, 2020-11
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    99
    Thermo Fisher paraformaldehyde pfa
    Paraformaldehyde Pfa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3743 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Thermo Fisher
    Average 99 stars, based on 3743 article reviews
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    91
    FUJIFILM paraformaldehyde phosphate buffer solution
    Paraformaldehyde Phosphate Buffer Solution, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 91/100, based on 328 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde phosphate buffer solution/product/FUJIFILM
    Average 91 stars, based on 328 article reviews
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    92
    Carl Roth GmbH pfa
    Loss of β1 integrin activity in brown adipocytes increases vascular permeability in <t>BAT</t> ( A ) Photograph of brown adipose tissue (BAT) from Kindlin-2 flox (Kind2 fl/fl) and Kindlin-2 flox; adiponectin-cre (Kind2 adipo-cre ) mice fed CD for 8 weeks and integrin beta 1 flox mice (Itgb1 fl/fl) and Itgb1 flox; adiponectin-cre (Itgb1 adipo-cre ) mice fed CD for 8 weeks after Evans blue (0.05 % in PBS) perfusion and fixation with <t>PFA</t> (4 %). ( B ) Representative images from B AT cryo-sections (30 µm) after Evans blue perfusion and PFA fixation from Kind2 fl/fl and Kind2 adipo-cre and Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD for 8 weeks (scale bar represents 100 µm). ( C ) Representative images from BAT of Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks after lectin perfusion and fixation with 4 % PFA. Lectin is represented in white color and F-actin in red (scale bar represents 25 µm). ( D ) Representative electron microscopy (EM) images showing adjacent parts of adipocytes in BAT from Kind2 fl/fl (WT) and Kind2 adipo-cre (KO) mice fed CD for 8 weeks (scale bar represents 1 µm). ( E ) Basal lamina thickness quantification (µm) measured by EM at 15 different points per vessels. Mean value is calculated per vessel and mouse in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( F ) Endothelial vesicle density (per µm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( G ) Endothelial overlap (nm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( H ) Representative western blot images for UCP1 and α-tubulin in BAT from Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks (n=5-4). ( I ) Representative H E staining of BAT sections from Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD and HFD for 14 weeks (scale bar represents 100 µm). ( J ) Body-weight (g) correlated to energy expenditure (kcal/h) in Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks during acute cold exposure at 4 °C for 5 hours (n=5-4). Data are shown as mean ± SEM. Statistics were calculated using two-way ANOVA with Tukey’s multiple comparison post-hoc test (**p
    Pfa, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 92/100, based on 388 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa/product/Carl Roth GmbH
    Average 92 stars, based on 388 article reviews
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    pfa - by Bioz Stars, 2020-11
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    93
    Merck & Co pfa
    Loss of β1 integrin activity in brown adipocytes increases vascular permeability in <t>BAT</t> ( A ) Photograph of brown adipose tissue (BAT) from Kindlin-2 flox (Kind2 fl/fl) and Kindlin-2 flox; adiponectin-cre (Kind2 adipo-cre ) mice fed CD for 8 weeks and integrin beta 1 flox mice (Itgb1 fl/fl) and Itgb1 flox; adiponectin-cre (Itgb1 adipo-cre ) mice fed CD for 8 weeks after Evans blue (0.05 % in PBS) perfusion and fixation with <t>PFA</t> (4 %). ( B ) Representative images from B AT cryo-sections (30 µm) after Evans blue perfusion and PFA fixation from Kind2 fl/fl and Kind2 adipo-cre and Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD for 8 weeks (scale bar represents 100 µm). ( C ) Representative images from BAT of Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks after lectin perfusion and fixation with 4 % PFA. Lectin is represented in white color and F-actin in red (scale bar represents 25 µm). ( D ) Representative electron microscopy (EM) images showing adjacent parts of adipocytes in BAT from Kind2 fl/fl (WT) and Kind2 adipo-cre (KO) mice fed CD for 8 weeks (scale bar represents 1 µm). ( E ) Basal lamina thickness quantification (µm) measured by EM at 15 different points per vessels. Mean value is calculated per vessel and mouse in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( F ) Endothelial vesicle density (per µm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( G ) Endothelial overlap (nm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( H ) Representative western blot images for UCP1 and α-tubulin in BAT from Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks (n=5-4). ( I ) Representative H E staining of BAT sections from Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD and HFD for 14 weeks (scale bar represents 100 µm). ( J ) Body-weight (g) correlated to energy expenditure (kcal/h) in Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks during acute cold exposure at 4 °C for 5 hours (n=5-4). Data are shown as mean ± SEM. Statistics were calculated using two-way ANOVA with Tukey’s multiple comparison post-hoc test (**p
    Pfa, supplied by Merck & Co, used in various techniques. Bioz Stars score: 93/100, based on 407 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa/product/Merck & Co
    Average 93 stars, based on 407 article reviews
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    pfa - by Bioz Stars, 2020-11
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    93
    Santa Cruz Biotechnology pfa
    Western blot assay (a) and immunofluorescence (b) experiments showing nAG protein expression in primary human fibroblasts. (a) Western blot was performed after 24 h and 48 h of transfection. The cells were lysed by using RIPA cocktail; proteins were separated on 12% SDS polyacrylamide gel. The primary antibody V5 probe and HRP-conjugated secondary antibody were used for nAG protein detection in the following lanes: nontransfected fibroblasts (negative control 1), fibroblasts with nAG plasmid without lipofection (negative control 2), nAG transfected fibroblasts tested after 24 h and nAG transfected fibroblasts tested after 48 h. (b) Immunofluorescence assay was performed after 48 h of transfection and cells were fixed and <t>permeabilized</t> by 2% <t>PFA/0.1%</t> Triton x-100. V5 probe was used as primary antibody and green fluorescence was detected by using FITC-conjugated secondary antibody. Compare nAG protein expression in nontransfected fibroblasts (negative control) to nAG expression in transfected fibroblasts (magnification 40x).
    Pfa, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1451 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1451 article reviews
    Price from $9.99 to $1999.99
    pfa - by Bioz Stars, 2020-11
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    90
    Dade Behring pfa 100
    NAC increased the closure time measured by <t>PFA-100.</t> Citrated whole blood was incubated with NAC at the indicated concentrations before being perfused through either a collagen/ADP cartridge ( A ) or a collagen/epinephrine cartridge ( B ) in the PFA-100 device. The closure times increased as the concentration of NAC increased. The machine stopped automatically if closure times were longer than 300 seconds. The mean difference (± SEM) in closure times measured at 10 mM NAC and at 2 mM were significant in both cartridges ( A : 140 ± 65.6 seconds, P
    Pfa 100, supplied by Dade Behring, used in various techniques. Bioz Stars score: 90/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa 100/product/Dade Behring
    Average 90 stars, based on 72 article reviews
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    pfa 100 - by Bioz Stars, 2020-11
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    99
    Millipore paraformaldehyde pfa solution
    NAC increased the closure time measured by <t>PFA-100.</t> Citrated whole blood was incubated with NAC at the indicated concentrations before being perfused through either a collagen/ADP cartridge ( A ) or a collagen/epinephrine cartridge ( B ) in the PFA-100 device. The closure times increased as the concentration of NAC increased. The machine stopped automatically if closure times were longer than 300 seconds. The mean difference (± SEM) in closure times measured at 10 mM NAC and at 2 mM were significant in both cartridges ( A : 140 ± 65.6 seconds, P
    Paraformaldehyde Pfa Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 159 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa solution/product/Millipore
    Average 99 stars, based on 159 article reviews
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    paraformaldehyde pfa solution - by Bioz Stars, 2020-11
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    90
    Siemens Healthineers pfa 100
    Blockage of extracellular CyPA through 8H7-mAb does not affect hemostasis (A/B) 8H7-mAb treated mice did not exhibita prolonged tail bleeding time (A). After tail bleeding time the blood was collected and the number of platelets (B) and leukocytes (C) were estimated for both groups. (D) An in vivo treatment with 8H7-mAb did not change the surface expression of CD42b, CD29, CD41, CD49b, GPVI or the active form of α IIb β 3 on platelets compared to IgG control treated mice. (F/G) Every dot shows the response of one donor for the prothrombin time (E) and the activated partial thromboplastin time (F) (n=8). (G) <t>PFA-100</t> measurement was performed with human blood. With the cartilages P 2 Y, Collagen/Epinephrine and Collage/ADP no difference could be detected.
    Pfa 100, supplied by Siemens Healthineers, used in various techniques. Bioz Stars score: 90/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa 100/product/Siemens Healthineers
    Average 90 stars, based on 61 article reviews
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    pfa 100 - by Bioz Stars, 2020-11
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    93
    Merck KGaA pfa
    Blockage of extracellular CyPA through 8H7-mAb does not affect hemostasis (A/B) 8H7-mAb treated mice did not exhibita prolonged tail bleeding time (A). After tail bleeding time the blood was collected and the number of platelets (B) and leukocytes (C) were estimated for both groups. (D) An in vivo treatment with 8H7-mAb did not change the surface expression of CD42b, CD29, CD41, CD49b, GPVI or the active form of α IIb β 3 on platelets compared to IgG control treated mice. (F/G) Every dot shows the response of one donor for the prothrombin time (E) and the activated partial thromboplastin time (F) (n=8). (G) <t>PFA-100</t> measurement was performed with human blood. With the cartilages P 2 Y, Collagen/Epinephrine and Collage/ADP no difference could be detected.
    Pfa, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 447 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa/product/Merck KGaA
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    pfa - by Bioz Stars, 2020-11
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    92
    Merck KGaA paraformaldehyde pfa
    Protocol for 3D blood vessel visualization in whole mouse brains. (a) Sample preparation: During animal sacrifice mice are transcardially perfused with 40 mL of phosphate-buffered saline <t>(PBS)</t> containing 100 U/mL heparin, followed by 20 mL of 4% paraformaldehyde <t>(PFA)</t> in PBS and 10 mL hydrogel composed of 2% gelatin and 0.1% FITC-conjugated albumin in PBS. (b) Whole brain clearing: In the optimized protocol, the brain specimen is incubated in each clearing solution (tetrahydrofuran (THF) used for dehydration and lipid solvation, dibenzyl ether (DBE) for matching of refractive index) for 12 h at room temperature with constant shaking in dark bottles. To ensure complete dehydration, the 100% THF step was performed twice. (c) Light sheet microscopy: Cleared brains immersed in DBE were imaged in a dorso-ventral direction using either 1.6 × or 6.4 × magnification and corresponding step sizes of 6 µm and 4 µm in the LaVision Biotec ultramicroscope.
    Paraformaldehyde Pfa, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 92/100, based on 362 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Merck KGaA
    Average 92 stars, based on 362 article reviews
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    paraformaldehyde pfa - by Bioz Stars, 2020-11
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    91
    Siemens AG pfa 100
    Platelet activation through mechanosensation and relative rate of energy consumption. GPIb? initially tethers VWF attached to exposed collagen. GP VI binds exposed collagen leading to activation of Syk and PLC?, increasing levels of cytoplasmic calcium. Soluble agonists act via g-protein coupled receptors to increase cytosolic calcium – enhancing expression of GP IIb-IIIa through TRAP1 and talin dependent pathways. Platelet pseudopodia are driven by actin–myosin interactions and influenced by shear deceleration as platelet velocity decreases after adhesion. Pseudopodia restructuring during aggregation stabilizes clot formation. Forces transmitted through GP IV and GP IIb-IIIa upregulates Src family kinases leading to myosin activation and platelet contraction via a ROCK and calcium dependent pathway. Energy consumption increases during aggregation and secretion. Devices exist to measure platelet function along the activation pathway. The XF 27–3 is able to continually measure oxygen consumption. FDA approved devices like the <t>PFA-100,</t> MultiPlate, Chrono-log and VerifyNow measure adhesion and aggregation. TEG-PM and ROTEM calculate platelet contribution to clot strength. Experimental devices measuring contraction include the Hemodyne, micro-posts and contraction cytometers.
    Pfa 100, supplied by Siemens AG, used in various techniques. Bioz Stars score: 91/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa 100/product/Siemens AG
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    99
    Thermo Fisher methanol free paraformaldehyde
    Platelet activation through mechanosensation and relative rate of energy consumption. GPIb? initially tethers VWF attached to exposed collagen. GP VI binds exposed collagen leading to activation of Syk and PLC?, increasing levels of cytoplasmic calcium. Soluble agonists act via g-protein coupled receptors to increase cytosolic calcium – enhancing expression of GP IIb-IIIa through TRAP1 and talin dependent pathways. Platelet pseudopodia are driven by actin–myosin interactions and influenced by shear deceleration as platelet velocity decreases after adhesion. Pseudopodia restructuring during aggregation stabilizes clot formation. Forces transmitted through GP IV and GP IIb-IIIa upregulates Src family kinases leading to myosin activation and platelet contraction via a ROCK and calcium dependent pathway. Energy consumption increases during aggregation and secretion. Devices exist to measure platelet function along the activation pathway. The XF 27–3 is able to continually measure oxygen consumption. FDA approved devices like the <t>PFA-100,</t> MultiPlate, Chrono-log and VerifyNow measure adhesion and aggregation. TEG-PM and ROTEM calculate platelet contribution to clot strength. Experimental devices measuring contraction include the Hemodyne, micro-posts and contraction cytometers.
    Methanol Free Paraformaldehyde, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 157 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Fisher Scientific pfa
    Platelet activation through mechanosensation and relative rate of energy consumption. GPIb? initially tethers VWF attached to exposed collagen. GP VI binds exposed collagen leading to activation of Syk and PLC?, increasing levels of cytoplasmic calcium. Soluble agonists act via g-protein coupled receptors to increase cytosolic calcium – enhancing expression of GP IIb-IIIa through TRAP1 and talin dependent pathways. Platelet pseudopodia are driven by actin–myosin interactions and influenced by shear deceleration as platelet velocity decreases after adhesion. Pseudopodia restructuring during aggregation stabilizes clot formation. Forces transmitted through GP IV and GP IIb-IIIa upregulates Src family kinases leading to myosin activation and platelet contraction via a ROCK and calcium dependent pathway. Energy consumption increases during aggregation and secretion. Devices exist to measure platelet function along the activation pathway. The XF 27–3 is able to continually measure oxygen consumption. FDA approved devices like the <t>PFA-100,</t> MultiPlate, Chrono-log and VerifyNow measure adhesion and aggregation. TEG-PM and ROTEM calculate platelet contribution to clot strength. Experimental devices measuring contraction include the Hemodyne, micro-posts and contraction cytometers.
    Pfa, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 456 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa/product/Fisher Scientific
    Average 92 stars, based on 456 article reviews
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    pfa - by Bioz Stars, 2020-11
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    pfa  (Avantor)
    92
    Avantor pfa
    Platelet activation through mechanosensation and relative rate of energy consumption. GPIb? initially tethers VWF attached to exposed collagen. GP VI binds exposed collagen leading to activation of Syk and PLC?, increasing levels of cytoplasmic calcium. Soluble agonists act via g-protein coupled receptors to increase cytosolic calcium – enhancing expression of GP IIb-IIIa through TRAP1 and talin dependent pathways. Platelet pseudopodia are driven by actin–myosin interactions and influenced by shear deceleration as platelet velocity decreases after adhesion. Pseudopodia restructuring during aggregation stabilizes clot formation. Forces transmitted through GP IV and GP IIb-IIIa upregulates Src family kinases leading to myosin activation and platelet contraction via a ROCK and calcium dependent pathway. Energy consumption increases during aggregation and secretion. Devices exist to measure platelet function along the activation pathway. The XF 27–3 is able to continually measure oxygen consumption. FDA approved devices like the <t>PFA-100,</t> MultiPlate, Chrono-log and VerifyNow measure adhesion and aggregation. TEG-PM and ROTEM calculate platelet contribution to clot strength. Experimental devices measuring contraction include the Hemodyne, micro-posts and contraction cytometers.
    Pfa, supplied by Avantor, used in various techniques. Bioz Stars score: 92/100, based on 455 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfa/product/Avantor
    Average 92 stars, based on 455 article reviews
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    pfa - by Bioz Stars, 2020-11
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    Image Search Results


    Loss of β1 integrin activity in brown adipocytes increases vascular permeability in BAT ( A ) Photograph of brown adipose tissue (BAT) from Kindlin-2 flox (Kind2 fl/fl) and Kindlin-2 flox; adiponectin-cre (Kind2 adipo-cre ) mice fed CD for 8 weeks and integrin beta 1 flox mice (Itgb1 fl/fl) and Itgb1 flox; adiponectin-cre (Itgb1 adipo-cre ) mice fed CD for 8 weeks after Evans blue (0.05 % in PBS) perfusion and fixation with PFA (4 %). ( B ) Representative images from B AT cryo-sections (30 µm) after Evans blue perfusion and PFA fixation from Kind2 fl/fl and Kind2 adipo-cre and Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD for 8 weeks (scale bar represents 100 µm). ( C ) Representative images from BAT of Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks after lectin perfusion and fixation with 4 % PFA. Lectin is represented in white color and F-actin in red (scale bar represents 25 µm). ( D ) Representative electron microscopy (EM) images showing adjacent parts of adipocytes in BAT from Kind2 fl/fl (WT) and Kind2 adipo-cre (KO) mice fed CD for 8 weeks (scale bar represents 1 µm). ( E ) Basal lamina thickness quantification (µm) measured by EM at 15 different points per vessels. Mean value is calculated per vessel and mouse in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( F ) Endothelial vesicle density (per µm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( G ) Endothelial overlap (nm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( H ) Representative western blot images for UCP1 and α-tubulin in BAT from Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks (n=5-4). ( I ) Representative H E staining of BAT sections from Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD and HFD for 14 weeks (scale bar represents 100 µm). ( J ) Body-weight (g) correlated to energy expenditure (kcal/h) in Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks during acute cold exposure at 4 °C for 5 hours (n=5-4). Data are shown as mean ± SEM. Statistics were calculated using two-way ANOVA with Tukey’s multiple comparison post-hoc test (**p

    Journal: bioRxiv

    Article Title: Active integrins regulate white adipose tissue insulin sensitivity and brown fat thermogenesis

    doi: 10.1101/2020.06.29.178020

    Figure Lengend Snippet: Loss of β1 integrin activity in brown adipocytes increases vascular permeability in BAT ( A ) Photograph of brown adipose tissue (BAT) from Kindlin-2 flox (Kind2 fl/fl) and Kindlin-2 flox; adiponectin-cre (Kind2 adipo-cre ) mice fed CD for 8 weeks and integrin beta 1 flox mice (Itgb1 fl/fl) and Itgb1 flox; adiponectin-cre (Itgb1 adipo-cre ) mice fed CD for 8 weeks after Evans blue (0.05 % in PBS) perfusion and fixation with PFA (4 %). ( B ) Representative images from B AT cryo-sections (30 µm) after Evans blue perfusion and PFA fixation from Kind2 fl/fl and Kind2 adipo-cre and Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD for 8 weeks (scale bar represents 100 µm). ( C ) Representative images from BAT of Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks after lectin perfusion and fixation with 4 % PFA. Lectin is represented in white color and F-actin in red (scale bar represents 25 µm). ( D ) Representative electron microscopy (EM) images showing adjacent parts of adipocytes in BAT from Kind2 fl/fl (WT) and Kind2 adipo-cre (KO) mice fed CD for 8 weeks (scale bar represents 1 µm). ( E ) Basal lamina thickness quantification (µm) measured by EM at 15 different points per vessels. Mean value is calculated per vessel and mouse in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( F ) Endothelial vesicle density (per µm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( G ) Endothelial overlap (nm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( H ) Representative western blot images for UCP1 and α-tubulin in BAT from Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks (n=5-4). ( I ) Representative H E staining of BAT sections from Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD and HFD for 14 weeks (scale bar represents 100 µm). ( J ) Body-weight (g) correlated to energy expenditure (kcal/h) in Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks during acute cold exposure at 4 °C for 5 hours (n=5-4). Data are shown as mean ± SEM. Statistics were calculated using two-way ANOVA with Tukey’s multiple comparison post-hoc test (**p

    Article Snippet: Histology and imagingSCF, PGF, BAT, and liver were taken and fixed in 4% PFA (Carl Roth, Germany) at room temperature for 24 h dehydrated in an ascending row of ethanol (70%-100%) and xylene.

    Techniques: Activity Assay, Permeability, Mouse Assay, Electron Microscopy, Western Blot, Staining

    ( A ) Photograph of subcutaneous (SCF) and perigonadal (PGF) adipose tissue from Kindlin-2 flox (Kind2 fl/fl) and Kindlin-2 flox; adiponectin-cre (Kind2 adipo-cre ) mice fed CD for 8 weeks after Evans blue (0.05 % in PBS) perfusion and fixation with 4 % PFA. ( B ) Photograph of SCF and PGF adipose tissue from integrin β1 flox mice (Itgb1 fl/fl) and Itgb1 flox; adiponectin-cre (Itgb1 adipo-cre ) mice fed CD for 8 weeks after Evans blue (0.05 % in PBS) perfusion and fixation with 4 % PFA. ( C ) Values distribution of basal lamina thickness quantification (µm) measured by EM at 15 different points to calculate the mean value per vessel and mouse in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( D ) Frequency distribution of endothelial vesicle density (per µm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( E ) Relative gene expression of Itgb1 and Itgb3 in brown adipose tissue (BAT) from Kind2 fl/fl and Kind2 adipo-cre mice fed HFD for 14 weeks (n=5-4). ( F ) Relative gene expression of Ucp1 in BAT from Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD (n=6-5) and HFD for 14 weeks (n=5-4). ( G ) Food intake (g) per day in Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks (n=5-4). ( H ) Respiratory quotient (RER) average of 3 days in Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD (n=5-4) for 14 weeks during acute cold exposure at 4 °C for 5 hours (n=5-4). Data are shown as mean ± SEM. Statistics were calculated using two-way ANOVA with Tukey’s multiple comparison post-hoc test (** P

    Journal: bioRxiv

    Article Title: Active integrins regulate white adipose tissue insulin sensitivity and brown fat thermogenesis

    doi: 10.1101/2020.06.29.178020

    Figure Lengend Snippet: ( A ) Photograph of subcutaneous (SCF) and perigonadal (PGF) adipose tissue from Kindlin-2 flox (Kind2 fl/fl) and Kindlin-2 flox; adiponectin-cre (Kind2 adipo-cre ) mice fed CD for 8 weeks after Evans blue (0.05 % in PBS) perfusion and fixation with 4 % PFA. ( B ) Photograph of SCF and PGF adipose tissue from integrin β1 flox mice (Itgb1 fl/fl) and Itgb1 flox; adiponectin-cre (Itgb1 adipo-cre ) mice fed CD for 8 weeks after Evans blue (0.05 % in PBS) perfusion and fixation with 4 % PFA. ( C ) Values distribution of basal lamina thickness quantification (µm) measured by EM at 15 different points to calculate the mean value per vessel and mouse in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( D ) Frequency distribution of endothelial vesicle density (per µm) measured by EM in BAT from Kind2 fl/fl and Kind2 adipo-cre mice fed CD for 8 weeks (n=3). ( E ) Relative gene expression of Itgb1 and Itgb3 in brown adipose tissue (BAT) from Kind2 fl/fl and Kind2 adipo-cre mice fed HFD for 14 weeks (n=5-4). ( F ) Relative gene expression of Ucp1 in BAT from Itgb1 fl/fl and Itgb1 adipo-cre mice fed CD (n=6-5) and HFD for 14 weeks (n=5-4). ( G ) Food intake (g) per day in Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD for 14 weeks (n=5-4). ( H ) Respiratory quotient (RER) average of 3 days in Itgb1 fl/fl and Itgb1 adipo-cre mice fed HFD (n=5-4) for 14 weeks during acute cold exposure at 4 °C for 5 hours (n=5-4). Data are shown as mean ± SEM. Statistics were calculated using two-way ANOVA with Tukey’s multiple comparison post-hoc test (** P

    Article Snippet: Histology and imagingSCF, PGF, BAT, and liver were taken and fixed in 4% PFA (Carl Roth, Germany) at room temperature for 24 h dehydrated in an ascending row of ethanol (70%-100%) and xylene.

    Techniques: Mouse Assay, Expressing

    Western blot assay (a) and immunofluorescence (b) experiments showing nAG protein expression in primary human fibroblasts. (a) Western blot was performed after 24 h and 48 h of transfection. The cells were lysed by using RIPA cocktail; proteins were separated on 12% SDS polyacrylamide gel. The primary antibody V5 probe and HRP-conjugated secondary antibody were used for nAG protein detection in the following lanes: nontransfected fibroblasts (negative control 1), fibroblasts with nAG plasmid without lipofection (negative control 2), nAG transfected fibroblasts tested after 24 h and nAG transfected fibroblasts tested after 48 h. (b) Immunofluorescence assay was performed after 48 h of transfection and cells were fixed and permeabilized by 2% PFA/0.1% Triton x-100. V5 probe was used as primary antibody and green fluorescence was detected by using FITC-conjugated secondary antibody. Compare nAG protein expression in nontransfected fibroblasts (negative control) to nAG expression in transfected fibroblasts (magnification 40x).

    Journal: BioMed Research International

    Article Title: Salamander-Derived, Human-Optimized nAG Protein Suppresses Collagen Synthesis and Increases Collagen Degradation in Primary Human Fibroblasts

    doi: 10.1155/2013/384091

    Figure Lengend Snippet: Western blot assay (a) and immunofluorescence (b) experiments showing nAG protein expression in primary human fibroblasts. (a) Western blot was performed after 24 h and 48 h of transfection. The cells were lysed by using RIPA cocktail; proteins were separated on 12% SDS polyacrylamide gel. The primary antibody V5 probe and HRP-conjugated secondary antibody were used for nAG protein detection in the following lanes: nontransfected fibroblasts (negative control 1), fibroblasts with nAG plasmid without lipofection (negative control 2), nAG transfected fibroblasts tested after 24 h and nAG transfected fibroblasts tested after 48 h. (b) Immunofluorescence assay was performed after 48 h of transfection and cells were fixed and permeabilized by 2% PFA/0.1% Triton x-100. V5 probe was used as primary antibody and green fluorescence was detected by using FITC-conjugated secondary antibody. Compare nAG protein expression in nontransfected fibroblasts (negative control) to nAG expression in transfected fibroblasts (magnification 40x).

    Article Snippet: Immunofluorescence Transfected cells were plated at density of 8 × 104 cells/well, and after 48 hours, medium was removed and cells were fixed and permeabilized by 2% PFA (Santa Cruz)/0.1% Triton x-100 (LKB Bromma, Sweden) in PBS for 30 minutes.

    Techniques: Western Blot, Immunofluorescence, Expressing, Transfection, Negative Control, Plasmid Preparation, Fluorescence

    NAC increased the closure time measured by PFA-100. Citrated whole blood was incubated with NAC at the indicated concentrations before being perfused through either a collagen/ADP cartridge ( A ) or a collagen/epinephrine cartridge ( B ) in the PFA-100 device. The closure times increased as the concentration of NAC increased. The machine stopped automatically if closure times were longer than 300 seconds. The mean difference (± SEM) in closure times measured at 10 mM NAC and at 2 mM were significant in both cartridges ( A : 140 ± 65.6 seconds, P

    Journal: The Journal of Clinical Investigation

    Article Title: N-acetylcysteine reduces the size and activity of von Willebrand factor in human plasma and mice

    doi: 10.1172/JCI41062

    Figure Lengend Snippet: NAC increased the closure time measured by PFA-100. Citrated whole blood was incubated with NAC at the indicated concentrations before being perfused through either a collagen/ADP cartridge ( A ) or a collagen/epinephrine cartridge ( B ) in the PFA-100 device. The closure times increased as the concentration of NAC increased. The machine stopped automatically if closure times were longer than 300 seconds. The mean difference (± SEM) in closure times measured at 10 mM NAC and at 2 mM were significant in both cartridges ( A : 140 ± 65.6 seconds, P

    Article Snippet: The following materials (with their sources indicated in parentheses) were used in this study: NAC (Acetadote; Cumberland Pharmaceuticals Inc.); PFA cartridges and PFA-100 (Dade Behring); calcein AM (Invitrogen); histamine, calcium ionophore, NEM, DTT, and tribromoethanol (Sigma-Aldrich); NEM-d5 (Cambridge Isotope Laboratories); rabbit anti-human vWF antibody (HRP conjugated; Dako North America Inc.); SeaKem gold agarose (Cambrex Bio Science Rockland Inc.); fixed platelets, ADP, collagen (for platelet aggregation), and AggRAM platelet aggregometer (Helena Laboratories Corp.); bovine collagen (for the collagen-binding assay) (MP Biomedicals; LLC); pooled normal plasma in citrate (Precision BioLogic); ristocetin (American Biochemical and Pharmaceutical Ltd.); cone-and-plate viscometer (HAAKE Rheostress 1; Thermo Fisher Scientific); endoproteinase Asp-N (Roche); Finnigan LTQ linear ion trap mass spectrometer (Thermo Electron Corporation).

    Techniques: Incubation, Concentration Assay

    Blockage of extracellular CyPA through 8H7-mAb does not affect hemostasis (A/B) 8H7-mAb treated mice did not exhibita prolonged tail bleeding time (A). After tail bleeding time the blood was collected and the number of platelets (B) and leukocytes (C) were estimated for both groups. (D) An in vivo treatment with 8H7-mAb did not change the surface expression of CD42b, CD29, CD41, CD49b, GPVI or the active form of α IIb β 3 on platelets compared to IgG control treated mice. (F/G) Every dot shows the response of one donor for the prothrombin time (E) and the activated partial thromboplastin time (F) (n=8). (G) PFA-100 measurement was performed with human blood. With the cartilages P 2 Y, Collagen/Epinephrine and Collage/ADP no difference could be detected.

    Journal: Thrombosis and haemostasis

    Article Title: Extracellular cyclophilin A augments platelet-dependent thrombosis and thrombo-inflammation

    doi: 10.1160/TH17-01-0067

    Figure Lengend Snippet: Blockage of extracellular CyPA through 8H7-mAb does not affect hemostasis (A/B) 8H7-mAb treated mice did not exhibita prolonged tail bleeding time (A). After tail bleeding time the blood was collected and the number of platelets (B) and leukocytes (C) were estimated for both groups. (D) An in vivo treatment with 8H7-mAb did not change the surface expression of CD42b, CD29, CD41, CD49b, GPVI or the active form of α IIb β 3 on platelets compared to IgG control treated mice. (F/G) Every dot shows the response of one donor for the prothrombin time (E) and the activated partial thromboplastin time (F) (n=8). (G) PFA-100 measurement was performed with human blood. With the cartilages P 2 Y, Collagen/Epinephrine and Collage/ADP no difference could be detected.

    Article Snippet: The influence of 8H7-mAb on platelets function was analyzed in a PFA-100 (Platelet Function Analyzer-100) system (Siemens Healthineers USA, Malvern, PA, USA).

    Techniques: Mouse Assay, In Vivo, Expressing

    Protocol for 3D blood vessel visualization in whole mouse brains. (a) Sample preparation: During animal sacrifice mice are transcardially perfused with 40 mL of phosphate-buffered saline (PBS) containing 100 U/mL heparin, followed by 20 mL of 4% paraformaldehyde (PFA) in PBS and 10 mL hydrogel composed of 2% gelatin and 0.1% FITC-conjugated albumin in PBS. (b) Whole brain clearing: In the optimized protocol, the brain specimen is incubated in each clearing solution (tetrahydrofuran (THF) used for dehydration and lipid solvation, dibenzyl ether (DBE) for matching of refractive index) for 12 h at room temperature with constant shaking in dark bottles. To ensure complete dehydration, the 100% THF step was performed twice. (c) Light sheet microscopy: Cleared brains immersed in DBE were imaged in a dorso-ventral direction using either 1.6 × or 6.4 × magnification and corresponding step sizes of 6 µm and 4 µm in the LaVision Biotec ultramicroscope.

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: 3D visualization and quantification of microvessels in the whole ischemic mouse brain using solvent-based clearing and light sheet microscopy

    doi: 10.1177/0271678X17698970

    Figure Lengend Snippet: Protocol for 3D blood vessel visualization in whole mouse brains. (a) Sample preparation: During animal sacrifice mice are transcardially perfused with 40 mL of phosphate-buffered saline (PBS) containing 100 U/mL heparin, followed by 20 mL of 4% paraformaldehyde (PFA) in PBS and 10 mL hydrogel composed of 2% gelatin and 0.1% FITC-conjugated albumin in PBS. (b) Whole brain clearing: In the optimized protocol, the brain specimen is incubated in each clearing solution (tetrahydrofuran (THF) used for dehydration and lipid solvation, dibenzyl ether (DBE) for matching of refractive index) for 12 h at room temperature with constant shaking in dark bottles. To ensure complete dehydration, the 100% THF step was performed twice. (c) Light sheet microscopy: Cleared brains immersed in DBE were imaged in a dorso-ventral direction using either 1.6 × or 6.4 × magnification and corresponding step sizes of 6 µm and 4 µm in the LaVision Biotec ultramicroscope.

    Article Snippet: Mice were deeply anesthetized by an intraperitoneal injection of 7% chloral hydrate and transcardially perfused with 40 mL of PBS containing 50 U/mL heparin (Ratiopharm, Ulm, Germany), followed by perfusion of 20 mL of 4% paraformaldehyde (PFA) (Merck-Millipore) in PBS.

    Techniques: Sample Prep, Mouse Assay, Incubation, Microscopy

    Platelet activation through mechanosensation and relative rate of energy consumption. GPIb? initially tethers VWF attached to exposed collagen. GP VI binds exposed collagen leading to activation of Syk and PLC?, increasing levels of cytoplasmic calcium. Soluble agonists act via g-protein coupled receptors to increase cytosolic calcium – enhancing expression of GP IIb-IIIa through TRAP1 and talin dependent pathways. Platelet pseudopodia are driven by actin–myosin interactions and influenced by shear deceleration as platelet velocity decreases after adhesion. Pseudopodia restructuring during aggregation stabilizes clot formation. Forces transmitted through GP IV and GP IIb-IIIa upregulates Src family kinases leading to myosin activation and platelet contraction via a ROCK and calcium dependent pathway. Energy consumption increases during aggregation and secretion. Devices exist to measure platelet function along the activation pathway. The XF 27–3 is able to continually measure oxygen consumption. FDA approved devices like the PFA-100, MultiPlate, Chrono-log and VerifyNow measure adhesion and aggregation. TEG-PM and ROTEM calculate platelet contribution to clot strength. Experimental devices measuring contraction include the Hemodyne, micro-posts and contraction cytometers.

    Journal: Platelets

    Article Title: Platelet biomechanics, platelet bioenergetics, and applications to clinical practice and translational research

    doi: 10.1080/09537104.2018.1453062

    Figure Lengend Snippet: Platelet activation through mechanosensation and relative rate of energy consumption. GPIb? initially tethers VWF attached to exposed collagen. GP VI binds exposed collagen leading to activation of Syk and PLC?, increasing levels of cytoplasmic calcium. Soluble agonists act via g-protein coupled receptors to increase cytosolic calcium – enhancing expression of GP IIb-IIIa through TRAP1 and talin dependent pathways. Platelet pseudopodia are driven by actin–myosin interactions and influenced by shear deceleration as platelet velocity decreases after adhesion. Pseudopodia restructuring during aggregation stabilizes clot formation. Forces transmitted through GP IV and GP IIb-IIIa upregulates Src family kinases leading to myosin activation and platelet contraction via a ROCK and calcium dependent pathway. Energy consumption increases during aggregation and secretion. Devices exist to measure platelet function along the activation pathway. The XF 27–3 is able to continually measure oxygen consumption. FDA approved devices like the PFA-100, MultiPlate, Chrono-log and VerifyNow measure adhesion and aggregation. TEG-PM and ROTEM calculate platelet contribution to clot strength. Experimental devices measuring contraction include the Hemodyne, micro-posts and contraction cytometers.

    Article Snippet: The PFA-100 (Siemens Corp., Washington D.C.) simulates in vitro shear stress and measures platelet adhesion and aggregation.

    Techniques: Activation Assay, Relative Rate, Planar Chromatography, Activated Clotting Time Assay, Expressing